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‘Synthesise,Characterise,Analyse(SCA)’:Amul8disciplinaryapproachtotacklingNew
Psychoac8veSubstances(NPS)
GavinMcLaughlin,B.Sc.,M.Sc.,MRSC.
PhDCandidate,AthloneIns2tuteofTechnology(AIT),IrelandResearchAssistant,TrinityCollegeDublin(TCD),Ireland
NewPsychoac8veSubstances(NPS)
• Inrecentyears,therehasbeenanunprecedentedincreaseinthe number, types and seizures of chemicals frequentlyreferredtoasnewpsychoac8vesubstances(NPS).
• The constant release of NPS onto the recrea8onal drugmarket con8nues to create challenges for scien8sts in theforensic,clinicalandtoxicologydisciplines.
24
41 49
74 81
101 100
0
20
40
60
80
100
120
2009 2010 2011 2012 2013 2014 2015
Num
ber o
f NPS
Year
Number of NPS detected
Categories
PhenazepamNitrozolamE8zolamChlorodiazepam
PentyloneDipentyloneDibutyloneEthylhexedroneN-ethylpentedroneα-PVPClephedroneMPHPPV8(α-PHP)PV9(α-POP)Chloro-PVP4-chloro-α-PVP
5F-MDMB-PINACA5F-AKB-485F-PB-22
3-Fluorophenmetrazine
25I-NBOMe
FentanylPentanoylfentanyl2-fluorofentanyl
MethyltryptamineDimethyltryptamine5-MeO-MiPT
NewdrugsencounteredbyForensicScienceIrelandin2016
(FSIAnnualReport2016)
Problems
• Mainproblems:– Absenceofchemicalandpharmacologicalinforma8on– Availability(online)– Availabilityofscien8ficandpatentliterature
• Forensic Science Laboratories tend to struggle when facedwiththechallengeofiden8fyingtheseNPS.
• Most laboratories do not have access to the sophis8catedanaly8cal techniques, suchasNMRandHR-MS, required fortheiden8fica8onofNPS.
• Absenceofchemicalreferencestandardsandsearchabledrugreferencelibraries.
CurrentabemptsforNPSiden8fica8on
• Examples:TheRESPONSEproject&WEDINOS
• Theseprojectsprovidesomeimportantanaly8caldetails
aboutNPS.
• However,theaimsoftheseprojectsarenottoprovideextensivechemicaldataorpharmacologicaldataonNPS.
ImportanceofOrganicSynthesis
• Facilitates the unambiguous identification of an unknown sample through targeted synthesis of all possible isomers.
• Allows synthesis of compounds, which may not be commerically available.
• Allows for the observation of synthesis related impurities or route specific by-products.
• Extending the synthesis of a currently emerging psychoactive substance to a range of analogues and derivatives provides important proactive analytical data to the forensic, law enforcement and clinical communities.
• Once synthesised, these compounds may be used for studies beyond analytical characterisation, such as exploration of pharmacological, metabolism or toxicological features.
Characterisa8on
Rangeofanaly8caltechniquesu8lisedtofacilitatethedetec8on,iden8fica8onandstructuralelucida8onofdrugs
ImportanceofPharmacologicalEvalua8on
• EvaluaGonofthemolecularmechanismofacGon– Techniqueused: In vitroMonoamine TransporterAssays in rat brain
synaptosomes.– Informa9on obtained: Informa8on on the re-uptake and releasing
proper8es of a selected drug at the dopamine transporters,norepinephrinetransportersandserotonintransportersaswellas itsselec8vityforandpotencyateachtransporter.
• EvaluaGonofthemetaboliteprofileofNPS– Techniqueused:pooledHumanLiverMicrosomesandLC-MS/MS– Informa9on obtained: Metabolite profile of a selected drug, which
aidsintheiden8fica8onofNPSinbiologicalmatrices.
Synthesis,characterisa8onandmonoaminetransporterac8vityoftheNewPsychoac8veSubstance(NPS)
Mexedroneanddifferen8a8onfromitsN-methoxyposi8onalisomer,N-methoxymephedrone
Background
NH2
O
HN
O
Mephedrone
β
α
Cathinone
N
O
α−PVP
GenericLegisla8on
NH2
O
β
α
Anysubstance(notbeingbupropion)structurallyderivedfrom2-amino-1-phenyl-1-propanonebymodifica8oninanyofthefollowingways:• Bysubs8tu8oninthephenylringtoanyextentwithalkyl,alkenyl,alkynyl,
alkoxy,alkylthio,alkylenedioxy,haloalkylorhalosubs8tuents,whetherorfurtherinthephenylringbyoneormoreotherunivalentsubs8tuents.
• Bysubs8tu8onatthe2-or3-posi8onofthepropanonesidechainwithalkylsubs8tuents
• Bysubs8tu8onatthenitrogenatomwithoneormorealkylordialkylgroups,orbyinclusionofnitrogenatominacyclicstructure
ReplacementforMephedrone
O HN
Mephedrone
O HN
Mexedrone
O
ON
O
N-Methoxymephedrone
A
OBBr
ON
OCH3NHOCH3 HCl
DIPEA(a) (b)
O
Cl
NaOMe
MeOH NaI
O
O
Br2
CH2Cl2
O
O
Br MeNH2O H
N
O(c) (d) (e) (f)
C
O
Cl(g)
DBr2
CH2Cl2
O
Cl
Br MeNH2O
Cl
HN
(h) (i)
O HN
Mephedrone
O HN
Mexedrone
O
ON
O
N-Methoxymephedrone
A
OBBr
ON
OCH3NHOCH3 HCl
DIPEA(a) (b)
O
Cl
NaOMe
MeOH NaI
O
O
Br2
CH2Cl2
O
O
Br MeNH2O H
N
O(c) (d) (e) (f)
C
O
Cl(g)
DBr2
CH2Cl2
O
Cl
Br MeNH2O
Cl
HN
(h) (i)
O HN
Mephedrone
O HN
Mexedrone
O
ON
O
N-Methoxymephedrone
A
OBBr
ON
OCH3NHOCH3 HCl
DIPEA(a) (b)
O
Cl
NaOMe
MeOH NaI
O
O
Br2
CH2Cl2
O
O
Br MeNH2O H
N
O(c) (d) (e) (f)
C
O
Cl(g)
DBr2
CH2Cl2
O
Cl
Br MeNH2O
Cl
HN
(h) (i)
SynthesisofMexedrone&N-methoxymephedrone
O
Cl
Sodium methoxide
O
OSodium iodide
Br2
O
O
Br
O
O
HN Methanolic methylamine
(3-chloro-1-(4-methyphenyl)propan-1-one) (3-Methoxy-1-(4-methylphenyl)propan-1-one)
2-Bromo-3-methoxy-1-(4-methyphenyl)propan-1-oneMexedrone
Br
O
NO
O
(alpha-bromo-4-methylpropiophenone) (N-Methoxymephedrone)
N,N-diisopropylethylamine
N,O-dimethylhydroxylamine. HCl
Mexedrone N-methoxymephedrone
Characterisa8on:GC-MS88
42
56
6573 105
119134 147
162175
40 60 80 100 120 140 160 180 200 m/z
280000
240000
200000
160000
120000
80000
40000
0
40 60 80 100 120 140 160 180 200 m/z
88
4256
6577 105
119
133 148 160 174 192
3000000
2600000
2200000
1800000
1400000
1000000
600000
200000
A
B
C
Mexedrone
N-Methoxymephedrone
m/z 207
207
207
M
m/z 207M
m/z 162 m/z 119 m/z 88 m/z 56 m/z 42
m/z 192 m/z 119 m/z 88 m/z 56 m/z 42
MexedroneEI-MS6.57 min
N-MethoxymephedroneEI-MS6.18 min
O HN
O
O HN O
HN
O
N NH
ON NH
ON
O
ON NO O
88
42
56
6573 105
119134 147
162175
40 60 80 100 120 140 160 180 200 m/z
280000
240000
200000
160000
120000
80000
40000
0
40 60 80 100 120 140 160 180 200 m/z
88
4256
6577 105
119
133 148 160 174 192
3000000
2600000
2200000
1800000
1400000
1000000
600000
200000
A
B
C
Mexedrone
N-Methoxymephedrone
m/z 207
207
207
M
m/z 207M
m/z 162 m/z 119 m/z 88 m/z 56 m/z 42
m/z 192 m/z 119 m/z 88 m/z 56 m/z 42
MexedroneEI-MS6.57 min
N-MethoxymephedroneEI-MS6.18 min
O HN
O
O HN O
HN
O
N NH
ON NH
ON
O
ON NO O
Characterisa8on:LC-MS
60 80 100 120 140 160 180 200 220 2400
20
40
60
80
100[%]
m/z
LC-ESI-Q-MSMexedrone8.01 min110 V
84.9
88.1 91
.1
105.
0 117.
111
9.1
120.
1
141.
014
9.1
148.
2
158.
116
0.2
163.
1
176.
117
7.2
190.
119
1.1
195.
9
208.
1
230.
2
[M + H]
[M + Na]
+
+
60 80 100 120 140 160 180 200 220 2400
20
40
60
80
100
m/z
LC-ESI-Q-MSN-Methoxymephedrone10.63 min110 V
208.
2
[M + H]+
85.0
91.1
105.
1
119.
1
140.
9
149.
0
176.
0
195.
9
120.
1
158.
116
1.1
7.5 10 12.5
0
20000
40000
60000
80000
100000
120000
140000
160000
8.01 min
10.63 min
min
A
B
60 80 100 120 140 160 180 200 220 2400
20
40
60
80
100[%]
m/z
LC-ESI-Q-MSMexedrone8.01 min110 V
84.9
88.1 91
.1
105.
0 117.
111
9.1
120.
1
141.
014
9.1
148.
2
158.
116
0.2
163.
1
176.
117
7.2
190.
119
1.1
195.
9
208.
1
230.
2
[M + H]
[M + Na]
+
+
60 80 100 120 140 160 180 200 220 2400
20
40
60
80
100
m/z
LC-ESI-Q-MSN-Methoxymephedrone10.63 min110 V
208.
2
[M + H]+
85.0
91.1
105.
1
119.
1
140.
9
149.
0
176.
0
195.
9
120.
1
158.
116
1.1
7.5 10 12.5
0
20000
40000
60000
80000
100000
120000
140000
160000
8.01 min
10.63 min
min
A
B
O
O
H2N
O H2N
O
O
HN
H
OH
O
HN
OH2
O
HN
O
HN
N
O
O
HN
HO
+
+ +Loss of MeOH
Observed: 208.13374C12H18NO2
+ : 208.13321Δ = 2.58 ppm
Observed: 176.10712C11H14NO+ : 176.10699Δ = 0.71 ppm
Observed: 190.12279C12H16NO+ : 190.12264Δ = 0.77 ppm
Loss of MeOH
Observed: 158.09650C11H12N+ : 158.09643Δ = 0.45 ppm
+ +
+
+
Observed: 119.04903C8H7O+ : 119.04914Δ = -0.90 ppm
Loss of CO
A
B
ON
O
ON
ON
H O
H+
O
Observed: 208.13297C12H18NO2+ : 208.13321
= -1.15 ppm
O
Observed:119.04901C8H7O+ : 119.04914
= -1.03 ppm
Loss of MeOH
Observed: 176.10689C11H14NO+ : 176.10699
= -0.60ppm
O
+H
HN
+
+ +
O
O
H2N
O H2N
O
O
HN
H
OH
O
HN
OH2
O
HN
O
HN
N
O
O
HN
HO
+
+ +Loss of MeOH
Observed: 208.13374C12H18NO2
+ : 208.13321Δ = 2.58 ppm
Observed: 176.10712C11H14NO+ : 176.10699Δ = 0.71 ppm
Observed: 190.12279C12H16NO+ : 190.12264Δ = 0.77 ppm
Loss of MeOH
Observed: 158.09650C11H12N+ : 158.09643Δ = 0.45 ppm
+ +
+
+
Observed: 119.04903C8H7O+ : 119.04914Δ = -0.90 ppm
Loss of CO
A
B
ON
O
ON
ON
H O
H+
O
Observed: 208.13297C12H18NO2+ : 208.13321
= -1.15 ppm
O
Observed:119.04901C8H7O+ : 119.04914
= -1.03 ppm
Loss of MeOH
Observed: 176.10689C11H14NO+ : 176.10699
= -0.60ppm
O
+H
HN
+
+ +
MonoamineTransporterAc8vityPharmacologysummary:• N-Methoxymephedroneshowedatransporter-meditatedreleasingprofile
comparabletomephedrone,althoughmuchlowerinpotency.
• Bycontrast,mexedronewasfoundtobeaweakmonoaminetransporteruptakeblockerandweakserotoninreleasingagent–‘hybrid’ac8vity.
• Thismightexplainwhythissubstancereceivedmixedreviewsonpsychonautforums.
Schema8crepresenta8onofthereleaseassaymethod
-9 -7 -5 -3
0
40
80
120
log [dose] M
% [3 H
]DA
upt
ake
-Chloromethylmephedrone
N-MethoxymephedroneMexedrone
Mephedrone
-9 -7 -5 -3
0
40
80
120
log [dose] M
% [3 H
]NE
upta
ke
-Chloromethylmephedrone
N-MethoxymephedroneMexedrone
Mephedrone
-9 -7 -5 -3
0
40
80
120
log [dose] M
% [3 H
]5H
T up
take
-Chloromethylmephedrone
N-MethoxymephedroneMexedrone
Mephedrone
-9 -7 -5 -3
0
40
80
120
log [dose] M
% [3 H
]MPP
+ re
leas
e
-Chloromethylmephedrone
N-MethoxymephedroneMexedrone
Mephedrone
DAT
-9 -7 -5 -3
0
40
80
120
log [dose] M
% [3 H
]MPP
+ re
leas
e
-Chloromethylmephedrone
N-MethoxymephedroneMexedrone
Mephedrone
NET
-9 -7 -5 -3
0
40
80
120
log [dose] M
% [3 H
]5H
T re
leas
e
-Chloromethylmephedrone
N-MethoxymephedroneMexedrone
Mephedrone
SERT
A
B
C
D
E
F
EffectsofMexedone,N-methoxymephedroneandmephedroneoninhibi8onofuptakeands8mula8onofreleaseatDAT,SERTandNETinratbrainsynaptosomes.
Mexedrone:Conclusion
• Chemicalprofileofmexedroneascertained.• Pharmacologicalevalua8onofmexedroneconcludeditis
muchlesspotentthanmephedrone.• Vendorsamplefoundtocontainmexedrone.• Thiswasthefirststudyonmexedrone,whichreiterates
thecon8nuousneedtoremainvigilantontheavailabilityofnewlyemergingpsychoac8vesubstances.
OverallConclusion
• This approach of combining chemistry withpharmacology allows for the genera8on of essen8aldata and is an effec8ve approach to tackling anincreasingly complex area of inves8ga8on wheregrowing demands are placed on inves8gators in thefieldofNPS.
OverallConclusion
• This ‘SCA’ approach has been used for several other NPSincluding:
4,4-dimethylaminorex
Mexedrone
O
O
HN
NO
NH2
NO
NH2
O
O
3’,4’-methylenedioxy-4-methylaminorex
N
Diphenidine
N
2-Methoxydiphenidine
O
AB-CHMFUPPYCA
F
NN
O NH
NH2
O
3-Fluorophenmetrazine
NHO
F
OO
F
4-Fluoromethylphenidate
Acknowledgements
• Dr.NoreenMorris(AthloneIns8tuteofTechnology)• Dr.PierceKavanaghandhiscolleagues(TrinityCollegeDublin)• Dr.SimonBrandt(LiverpoolJohnMooresUniversity)• Mr.JohnPower(ForensicScienceIreland)• Dr.GeraldineDowling(DublinIns8tuteofTechnology)• Dr.RachelChris8e(EMCDDA)• Dr.MichaelH.Baumannandhiscolleagues(Na8onalIns8tuteofDrug
Abuse)• Dr.VolkerAüwarterandhiscolleagues(UniversityMedicalCenter
Freiburg)
Publica8onlist1. GavinMcLaughlin,NoreenMorris,PierceV.Kavanagh,JohnD.Power,GeraldineDowling,BrendanTwamley,John
O’Brien,GaryHessman,BrianMurphy,DonnaWalther,JohnS.Par8lla,MichaelH.Baumann,SimonD.Brandt.Analy8calcharacteriza8onandpharmacologicalevalua8onofthenewpsychoac8vesubstance4-fluoromethylphenidate(4F-MPH)anddifferen8a8onbetweenthe(±)-threoand(±)-erythrodiastereomers.DrugTes8ngandAnalysis.2017,9,347-357.DOI:10.1002/dta.2167
2. JohnD.Power,PierceV.Kavanagh,GavinMcLaughlin,MichaelBarry,GeraldineDowling,SimonD.Brandt.‘APAANin
theneck’–Areflec8ononsomenovelimpuri8esfoundinseizedmaterialscontainingamphetamineinIrelandduringforensicanalysis.DrugTes8ngandAnalysis.2017.DOI:10.1002/dta.2194
3. GeraldineDowling,PierceV.Kavanagh,BrianTalbot,JohnO’Brien,GaryHessman,GavinMcLaughlin,BrendanTwamley,SimonD.Brandt.Outsmartedbynootropics?Aninves8ga8onintothethermaldegrada8onofmodafinil,modafinicacid,adrafinil,CRL-40,940andCRL-40,941intheGCinjector:forma8onof1,1,2,2-tetraphenylethaneanditstetrafluoroanalog.DrugTes8ngandAnalysis.2017,9,446-452.DOI:10.1002/dta.2142
4. GavinMcLaughlin,NoreenMorris,PierceV.Kavanagh,JohnD.Power,GeraldineDowling,BrendanTwamley,JohnO’Brien,BrianTalbot,DonnaWalther,JohnS.Par8lla,MichaelH.Baumann,SimonD.Brandt.Synthesis,characteriza8onandmonoaminetransporterac8vityofthenewpsychoac8vesubstancemexedroneanditsN-methoxyposi8onalisomer,N-methoxymephedrone.DrugTes8ngandAnalysis.2017,9,358-368.DOI:10.1002/dta.2053
5. FlorianFranz,VerenaAngerer,SimonD.Brandt,GavinMcLaughlin,PierceV.Kavanagh,BjoernMoosmann,VolkerAuwarter.Invitrometabolismofthesynthe8ccannabinoid3,5-AB-CHMFUPPYCAandits5,3-regioisomerandinves8ga8onoftheirthermalstability.DrugTes8ngandAnalysis.2017,9,311-316.DOI:10.1002/dta.1950.
6. GavinMcLaughlin,NoreenMorris,PierceV.Kavanagh,GeraldineDowling,JohnD.Power,BrendanTwamley,JohnO'Brien,BrianTalbot,HaraldH.Sibe,SimonD.Brandt.Testpurchase,synthesisandcharacteriza8onof3-fluorophenmetrazine(3-FPM)anddifferen8a8onfromitsortho-andpara-subs8tutedisomers.DrugTes8ngandAnalysis.2017,9,369-377.DOI:10.1002/dta.1945.
7. JohnD.Power,PierceV.Kavanagh,GavinMcLaughlin,GeraldineDowling,MichaelBarry,JohnO'Brien,BrianTalbot,BrendanTwamley,SimonD.Brandt.ForensicanalysisofP2Pderivedamphetaminesynthesisimpuri8es:iden8fica8onandcharacteriza8onofindeneby-products.DrugTes8ngandAnalysis.2017,9,446-452.DOI:10.1002/dta.1944.
8. GavinMcLaughlin,NoreenMorris,PierceV.Kavanagh,JohnD.Power,BrendanTwamley,JohnO’Brien,BrianTalbot,GeraldineDowling,SimonD.Brandt.Thesynthesisandcharacterisa8onofthe‘researchchemical’N-(1-amino-3-methyl-1-oxobutan-2-yl)-1-(cyclohexylmethyl)-3-(4-fluorophenyl)-1H-pyrazole-5-carboxamide(3,5-AB-CHMFUPPYCA)anddifferen8a8onfromits5,3-regioisomer.DrugTes8ngandAnalysis.2016,8,920-929.DOI:10.1002/dta.1864.
9. GavinMcLaughlin,NoreenMorris,PierceV.Kavanagh,JohnD.Power,JohnO’Brien,BrianTalbot,SimonP.Elliob,JasonWallach,HamiltonMorris,SimonD.Brandt.Testpurchase,synthesisandcharacteriza8onof2-methoxydiphenidine(MXP)anddifferen8a8onfromitsmeta-andpara-subs8tutedisomers.DrugTes8ngandAnalysis.2015,8,98-109.DOI:10.1002/dta.1800.
10. JohnD.Power,PierceKavanagh,GavinMcLaughlin,JohnO’Brien,BrianTalbot,MichaelBarry,BrendanTwamley,GeraldineDowling,SimonD.Brandt.Iden8fica8onandcharacteriza8onofanimidazoliumby-productformedduringthesynthesisof4-methylmethcathinone(mephedrone).DrugTes8ngandAnalysis.2015,7,894-902.DOI:10.1002/dta.1789.
11. GavinMcLaughlin,NoreenMorris,PierceV.Kavanagh,JohnD.Power,BrendanTwamley,JohnO’Brien,BrianTalbot,GeraldineDowling,OliviaMahony,SimonD.Brandt,JulianPatrick,RolandP.Archer,JohnS.Par8lla,MichaelH.Baumann.Synthesis,characteriza8onandmonoaminetransporterac8vityofthenewpsychoac8vesubstance3’,4’-methylenedioxy-4-methylaminorex(MDMAR).DrugTes8ngandAnalysis.2014,7,555-564.DOI:10.1002/dta.1732.
12. JasonWallach,PierceV.Kavanagh,GavinMcLaughlin,NoreenMorris,JohnD.Power,SimonP.Elliob,MarionS.Mercier,DavidLodge,HamiltonMorris,NicolaM.Dempster,SimonD.Brandt.Prepara8onandcharacteriza8onofthe‘researchchemical’diphenidine,itspyrrolidineanalogue,andtheir2,2-diphenylethylisomers.DrugTes8ngandAnalysis.2014,7,358-367.DOI:10.1002/dta.1689
Publica8onlist
